Bubble-particle dynamics in multiphase flow of capillary foams in a porous micromodel.

Surfactant-free capillary foams (CFs) are known to be remarkably tolerant to oil, and possess unique stability and flow properties. These properties result from the presence of oil-and-particle-coated bubbles that are interconnected by a dense particle-oil capillary network. In this work, we present a study of the dynamics of capillary foams flowing through a porous micromodel. We determine that despite the presence of oil-particle networks, CFs can flow through a microporous environment and that above a threshold flowrate, >80% of foam pumped through the micromodel can be recovered. In addition, we highlight the absence of steady state in CF flow and identify the underlying phenomena including the increasing apparent viscosity, reconfigurable flow paths, and intermittent clogging of the micromodel from an oil-particle composite and bubbles trapped in pores. We also characterize bubble dynamics and show that CFs surprisingly exhibit the same bubble generation and destruction mechanisms as classical foams despite the absence of surfactants. Our observations suggest that the porous medium plays a key role in generating uniformly sized bubbles and that capillary foams in a microporous environment tend to reconfigure their flow paths in a manner that may provide opportunities for increased sweep efficiency in enhanced oil recovery.

Single-molecule FISH in C. elegans embryos reveals early embryonic expression dynamics of par-2 , lgl-1 and chin-1 and possible differences between hyper-diverged strains.

Wild C. elegans strains harbor natural variation in developmental pathways, but investigating these differences requires precise and well-powered phenotyping methods. Here we employ a microfluidics platform for single-molecule FISH to simultaneously visualize the transcripts of three genes in embryos of two distinct strains. We capture transcripts at high resolution by developmental stage in over one hundred embryos of each strain and observe wide-scale conservation of expression, but subtle differences in par-2 and chin-1 abundance and rate of change. As both genes reside in a genomic interval of hyper-divergence, these results may reflect consequences of pathway evolution over long timescales.

Automated and Dynamic Control of Chemical Content in Droplets for Scalable Screens of Small Animals.

Screening functional phenotypes in small animals is important for genetics and drug discovery. Multiphase microfluidics has great potential for enhancing throughput but has been hampered by inefficient animal encapsulation and limited control over the animal's environment in droplets. Here, a highly efficient single-animal encapsulation unit, a liquid exchanger system for controlling the droplet chemical environment dynamically, and an automation scheme for the programming and robust execution of complex protocols are demonstrated. By careful use of interfacial forces, the liquid exchanger unit allows for adding and removing chemicals from a droplet and, therefore, generating chemical gradients inaccessible in previous multiphase systems. Using Caenorhabditis elegans as an example, it is demonstrated that these advances can serve to analyze dynamic phenotyping, such as behavior and neuronal activity, perform forward genetic screen, and are scalable to manipulate animals of different sizes. This platform paves the way for large-scale screens of complex dynamic phenotypes in small animals.

High-Temporal-Resolution smFISH Method for Gene Expression Studies in Caenorhabditis elegans Embryos.

Recent development in fluorescence-based molecular tools has contributed significantly to developmental studies, including embryogenesis. Many of these tools rely on multiple steps of sample manipulation, so obtaining large sample sizes presents a major challenge as it can be labor-intensive and time-consuming. However, large sample sizes are required to uncover critical aspects of embryogenesis, for example, subtle phenotypic differences or gene expression dynamics. This problem is particularly relevant for single-molecule fluorescence in situ hybridization (smFISH) studies in Caenorhabditis elegans embryogenesis. Microfluidics can help address this issue by allowing a large number of samples and parallelization of experiments. However, performing efficient reagent exchange on chip for large numbers of embryos remains a bottleneck. Here, we present a microfluidic pipeline for large-scale smFISH imaging of C. elegans embryos with minimized labor. We designed embryo traps and engineered a protocol allowing for efficient chemical exchange for hundreds of C. elegans embryos simultaneously. Furthermore, the device design and small footprint optimize imaging throughput by facilitating spatial registration and enabling minimal user input. We conducted the smFISH protocol on chip and demonstrated that image quality is preserved. With one device replacing the equivalent of 10 glass slides of embryos mounted manually, our microfluidic approach greatly increases throughput. Finally, to highlight the capability of our platform to perform longitudinal studies with high temporal resolution, we conducted a temporal analysis of par-1 gene expression in early C. elegans embryos. The method demonstrated here paves the way for systematic high-temporal-resolution studies that will benefit large-scale RNAi and drug screens and in systems beyond C. elegans embryos.

Hydrogel-droplet microfluidic platform for high-resolution imaging and sorting of early larval Caenorhabditis elegans.

The nematode Caenorhabditis elegans is an important model organism in research on neuroscience and development because of its stereotyped anatomy, relevance to human biology, and ease of culture and genetic manipulation. The first larval stage (L1) is of particular interest in many biological problems, including post-embryonic developmental processes and developmental decision-making, such as dauer formation. However, L1's small size and high mobility make it difficult to manipulate; particularly in microfluidic chips, which have been used to great advantage in handling larger larvae and adult animals, small features are difficult to fabricate and these structures often get clogged easily, making the devices less robust. We have developed a microfluidic device to overcome these challenges and enable high-resolution imaging and sorting of early larval stage C. elegans via encapsulation in droplets of a thermosensitive hydrogel. To achieve precise handling of early larval stage worms, we demonstrated on-chip production, storage, and sorting of hydrogel droplets. We also demonstrated temporary immobilization of the worms within the droplets, allowing high-resolution imaging with minimal physiological perturbations. Because of the ability to array hydrogel droplets for handling a large number of L1 worms in a robust way, we envision that this platform will be widely applicable to screening in various developmental studies.

A perspective on optical developments in microfluidic platforms for Caenorhabditis elegans research.

Microfluidics offers unique ways of handling and manipulating microorganisms, which has particularly benefited Caenorhabditis elegans research. Optics plays a major role in these microfluidic platforms, not only as a read-out for the biological systems of interest but also as a vehicle for applying perturbations to biological systems. Here, we describe different areas of research in C. elegans developmental biology and behavior neuroscience enabled by microfluidics combined with the optical components. In particular, we highlight the diversity of optical tools and methods in use and the strategies implemented in microfluidics to make the devices compatible with optical techniques. We also offer some thoughts on future challenges in adapting advancements in optics to microfluidic platforms.

Microfluidic-based generation of size-controlled, biofunctionalized synthetic polymer microgels for cell encapsulation.

Cell and islet microencapsulation in synthetic hydrogels provides an immunoprotective and cell-supportive microenvironment. A microfluidic strategy for the genaration of biofunctionalized, synthetic microgel particles with precise control over particle size and molecular permeability for cell and protein delivery is presented. These engineered capsules support high cell viability and function of encapsulated human stem cells and islets.